Disease Information:
Tuberculosis is an airborne disease that is spread from one person to another through the air. Symptoms of tuberculosis include coughing, chills,
loss of appetite, fever, and coughing blood. Even though the bacteria usually attacks the lungs, it also can affect the kidney spine, and brain.
It catalyzes the formation of S-adenosylmethionine (SAM) from Met and ATP. S-adenosylmethionine is an essential methyl donor in mycolipid biosynthesis. SAM synthetase is also involved in a subpathway involved in S-adenosyl-L-methionine biosyntheis which is part of the amino-acid biosynthesis pathway. Contains ideR protein in its genes which is a metal regulator and when it is in the presence of iron, it can regulate transcription.
Expression Information (has it been expressed in bacterial cells):
AVA0421 vector (derived from pET14b and provides cleavable 6xHis tags at the N-termini of the expressed proteins).
The recombinant plasmids were transformed into E. coli strain BL21(DE3)-R3-pRARE2 cells for expression testing.
Purification Method: Purified from 6xHis nickel-affinity tag
Image of protein (PyMol with features delineated and shown separately):
Target (protein/gene name):
S-adenosylmethionine synthetaseNCBI Gene # or RefSeq#:
Tuberculist: Rv1392NCBI Gene: 83332
GeneID: 886741
Link to PubChem:
https://www.ncbi.nlm.nih.gov/Structure/pdb/3TDE
Protein ID (NP or XP #) or Wolbachia#:
NP_215908Organism (including strain): Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Disease Information:Tuberculosis is an airborne disease that is spread from one person to another through the air. Symptoms of tuberculosis include coughing, chills,
loss of appetite, fever, and coughing blood. Even though the bacteria usually attacks the lungs, it also can affect the kidney spine, and brain.
Link to TDR Targets page (if present): http://tdrtargets.org/targets/view?gene_name=Rv1392
Link to Gene Database page (NCBI, Tuberculist , BioCyc, TBDB):
https://www.ncbi.nlm.nih.gov/gene/885638
Essentiality of this protein:
It catalyzes the formation of S-adenosylmethionine (SAM) from Met and ATP. S-adenosylmethionine is an essential methyl donor in mycolipid biosynthesis.SAM synthetase is also involved in a subpathway involved in S-adenosyl-L-methionine biosyntheis which is part of the amino-acid biosynthesis pathway.
Contains ideR protein in its genes which is a metal regulator and when it is in the presence of iron, it can regulate transcription.
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html):
This protein is a homotetramer.Complex of proteins?: No
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): 0.5 From TDR targets
EC#: 2.5.1.6
From UniPathway
Enzyme Assay information : Fluorometric pyrophosphate assay (blue fluorescence)
-- link to Sigma (or other company) page for assay (see Sigma links below)-- link (or citation) to paper that contains assay information: http://www.jst.go.jp/erato/asano/english/activity/AAM_Kameya_2013_2.pdf
-- links to assay reagents (substrates) pages. Pyrophosphate Assay Kit
List cost and quantity of substrate reagents, supplier, and catalog #
SIGMA:-Pyrophosphate assay kit (MAK168): $251.50
-Pyrophosphate reagent (P7275): $56.40 (1VL)
$282.50 (10VL)
Structure (PDB or Homology model): 3TDE
Current Inhibitors: (none for M. tuberulosis)Expression Information (has it been expressed in bacterial cells):
AVA0421 vector (derived from pET14b and provides cleavable 6xHis tags at the N-termini of the expressed proteins).The recombinant plasmids were transformed into E. coli strain BL21(DE3)-R3-pRARE2 cells for expression testing.
Purification Method: Purified from 6xHis nickel-affinity tag
Image of protein (PyMol with features delineated and shown separately):Amino Acid Sequence:
GPGSMVSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETL
VTTGQVHVVGEVTTSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVN
IGIGAQSPDIAQGVDTAHEARVEGAADPLDSQGAGDQGLMFGYAINATPEL
MPLPIALAHRLSRRLTEVRKNGVLPYLRPDGKTQVTIAYEDNVPVRLDTVV
ISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDASTVRVLVNPTGKFV
LGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMRW
VAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGE
VFDLRPGAIIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI
Length of your protein in Amino Acids: 261 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 43444.1
Molar Extinction coefficient of your protein at 280 nm wavelength
Ext. coefficient 30035Abs 0.1% (=1 g/l) 0.691, assuming all pairs of Cys residues form cystines
Ext. coefficient 29910
Abs 0.1% (=1 g/l) 0.688, assuming all Cys residues are reduced
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html):
CDS Gene:
ATGGTTTCTGAAAAAGGTCGTCTGTTTACGTCTGAGTCTGTTACGGAAGGCCATCCTGACAAAATTTGCGACGCTATCTCTGACTCTGTACTGGATGCACTCCTGGCGGCAGACCCTCGTTCTCGTGTTGCGGTTGAGACTC
TCGTAACCACCGGTCAGGTTCACGTTGTTGGTGAAGTTACCACCTCTGCAAAAGAAGCGTTTGCGGATATC
ACTAACACGGTCCGTGCGCGTATTCTGGAAATTGGCTACGACTCTTCTGACAAGGGTTTCGATGGTGCGAC
GTGCGGCGTGAACATCGGTATCGGTGCGCAGTCTCCTGATATTGCGCAGGGTGTTGACACCGCACACGAAG
CGCGTGTGGAAGGTGCTGCAGATCCTCTGGACTCTCAGGGTGCCGGTGACCAGGGCCTGATGTTCGGTTAC
GCAATCAATGCGACCCCTGAACTGATGCCGCTCCCGATCGCGCTGGCCCATCGCCTGTCTCGTCGTCTCAC
TGAGGTTCGTAAAAACGGTGTTCTCCCGTACCTCCGCCCAGATGGTAAGACCCAGGTTACTATCGCGTACG
AGGATAACGTTCCGGTTCGTCTCGACACTGTGGTAATCTCTACCCAGCACGCGGCTGACATCGACCTGGAG
AAAACCCTCGATCCGGACATCCGCGAGAAAGTTCTCAACACTGTCCTGGATGACCTCGCACATGAGACGCT
GGACGCATCCACCGTTCGTGTTCTGGTAAATCCTACGGGCAAATTCGTTCTGGGTGGTCCGATGGGTGATG
CGGGTCTGACTGGTCGCAAGATCATCGTAGACACCTACGGCGGTTGGGCCCGTCACGGTGGTGGTGCGTTC
AGCGGCAAAGACCCGTCTAAGGTTGACCGTTCTGCGGCCTATGCCATGCGTTGGGTTGCGAAAAACGTCGT
GGCAGCAGGTCTGGCGGAGCGTGTAGAAGTTCAAGTCGCGTATGCAATCGGTAAAGCGGCACCTGTTGGTC
TGTTCGTTGAAACTTTCGGCACCGAAACGGAAGATCCGGTAAAAATCGAAAAAGCGATTGGTGAGGTTTTT
GATCTGCGTCCGGGTGCCATCATTCGTGATCTGAATCTGCTCCGTCCAATCTACGCGCCGACCGCAGCGTA
CGGCCACTTTGGTCGTACCGACGTTGAACTGCCGTGGGAACAACTGGACAAGGTCGACGATCTCAAACGTG
CGATCTAA
GC% Content for gene: 55.2%
CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol:
1 ATGGTTTCTGAAAAAGGTCGTCTGTTTACGTCTGAGTC2 ATAGCGTCGCAAATTTTGTCAGGATGGCCTTCCGTAACAGACTCAGACGTAAACAGACGA
3 TGACAAAATTTGCGACGCTATCTCTGACTCTGTACTGGATGCACTCCTGGCGGCAGACCC
4 ACCTGACCGGTGGTTACGAGAGTCTCAACCGCAACACGAGAACGAGGGTCTGCCGCCAGG
5 CGTAACCACCGGTCAGGTTCACGTTGTTGGTGAAGTTACCACCTCTGCAAAAGAAGCGTT
6 TTCCAGAATACGCGCACGGACCGTGTTAGTGATATCCGCAAACGCTTCTTTTGCAGAGGT
7 CGTGCGCGTATTCTGGAAATTGGCTACGACTCTTCTGACAAGGGTTTCGATGGTGCGACG
8 AATATCAGGAGACTGCGCACCGATACCGATGTTCACGCCGCACGTCGCACCATCGAAACC
9 GTGCGCAGTCTCCTGATATTGCGCAGGGTGTTGACACCGCACACGAAGCGCGTGTGGAAG
10 CCCTGGTCACCGGCACCCTGAGAGTCCAGAGGATCTGCAGCACCTTCCACACGCGCTTCG
11 GTGCCGGTGACCAGGGCCTGATGTTCGGTTACGCAATCAATGCGACCCCTGAACTGATGC
12 TGAGACGACGAGACAGGCGATGGGCCAGCGCGATCGGGAGCGGCATCAGTTCAGGGGTCG
13 GCCTGTCTCGTCGTCTCACTGAGGTTCGTAAAAACGGTGTTCTCCCGTACCTCCGCCCAG
14 CGGAACGTTATCCTCGTACGCGATAGTAACCTGGGTCTTACCATCTGGGCGGAGGTACGG
15 CGTACGAGGATAACGTTCCGGTTCGTCTCGACACTGTGGTAATCTCTACCCAGCACGCGG
16 CTCGCGGATGTCCGGATCGAGGGTTTTCTCCAGGTCGATGTCAGCCGCGTGCTGGGTAGA
17 ATCCGGACATCCGCGAGAAAGTTCTCAACACTGTCCTGGATGACCTCGCACATGAGACGC
18 TTGCCCGTAGGATTTACCAGAACACGAACGGTGGATGCGTCCAGCGTCTCATGTGCGAGG
19 CTGGTAAATCCTACGGGCAAATTCGTTCTGGGTGGTCCGATGGGTGATGCGGGTCTGACT
20 ACGGGCCCAACCGCCGTAGGTGTCTACGATGATCTTGCGACCAGTCAGACCCGCATCACC
21 GCGGTTGGGCCCGTCACGGTGGTGGTGCGTTCAGCGGCAAAGACCCGTCTAAGGTTGACC
22 GACGTTTTTCGCAACCCAACGCATGGCATAGGCCGCAGAACGGTCAACCTTAGACGGGTC
23 GTTGGGTTGCGAAAAACGTCGTGGCAGCAGGTCTGGCGGAGCGTGTAGAAGTTCAAGTCG
24 CGAACAGACCAACAGGTGCCGCTTTACCGATTGCATACGCGACTTGAACTTCTACACGCT
25 GCACCTGTTGGTCTGTTCGTTGAAACTTTCGGCACCGAAACGGAAGATCCGGTAAAAATC
26 CCGGACGCAGATCAAAAACCTCACCAATCGCTTTTTCGATTTTTACCGGATCTTCCGTTT
27 GTTTTTGATCTGCGTCCGGGTGCCATCATTCGTGATCTGAATCTGCTCCGTCCAATCTAC
28 GTCGGTACGACCAAAGTGGCCGTACGCTGCGGTCGGCGCGTAGATTGGACGGAGCAGATT
29 CCACTTTGGTCGTACCGACGTTGAACTGCCGTGGGAACAACTGGACAAGGTCGACGATCT
30 TTAGATCGCACGTTTGAGATCGTCGACCTTGTCCA
GC% Content for gene (codon optimized): 55.7%
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
From Tuberculist (NCBI)
VSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVT
TSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARVEGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYLRPDG
KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST
VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR
WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA
IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI
Methionine added to beginning of sequence for E coli
MVSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVTTSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHE
ARVEGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYL
RPDGKTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHET
LDASTVRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRS
AAYAMRWVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEV
FDLRPGAIIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI
Primer design results for 'tail' primers (this is just 2 sequences):
Forward Tail Primer in 5' to 3' orientation:
5’ TACTTCCAATCCATGGTTTCTGAAAAAGGTCGT 3’
Reverse Tail Primer in 5' to 3' orientation:
5’ TATCCACCTTTACTGTTAGATCGCACGTTTGAGAT 3’
Range 1: 1 to 403Query_132989&tracks=[key:sequence_track,name:Sequence,display_name:Sequence,id:STD1,category:Sequence,annots:Sequence,ShowLabel:true][key:gene_model_track,CDSProductFeats:false][key:alignment_track,name:other%20alignments,annots:NG%20Alignments|Refseq%20Alignments|Gnomon%20Alignments|Unnamed,shown:false]&v=0:423&appname=ncbiblast&link_loc=fromHSP|Graphics
Alignment statistics for match #1||~ Score
Kalign of PDB, NCBI, and TubercuList amino acid sequences:
PDB Sequence:http://www.rcsb.org/pdb/explore/remediatedSequence.do?structureId=3tde
>3TDE:D|PDBID|CHAIN|SEQUENCE
GPGSMVSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVTTSAKEAFADITNTVR
ARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARVEGAADPLDSQGAGDQGLMFGYAINATPELMPLPIA
LAHRLSRRLTEVRKNGVLPYLRPDGKTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHET
LDASTVRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMRWVAKNVVAAGLAERV
EVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGAIIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDK
VDDLKRAI
NCBI Sequence: THIS WAS USED TO DESIGN PRIMER (SUMMER 2017)
https://www.ncbi.nlm.nih.gov/protein/15608531?report=fasta
TubercuList Sequence:
http://tuberculist.epfl.ch/quicksearch.php?gene+name=Rv1392&submit=Search
>M. tuberculosis H37Rv|Rv1392|metK
VSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVT
TSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARV
EGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYLRPDG
KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST
VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR
WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA
IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI
<span style="font-size: 11.869px;">Query 6 VSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVT 65 +SEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVT Sbjct 1 MSEKGRLFTSESVTEGHPDKICDAISDSVLDALLAADPRSRVAVETLVTTGQVHVVGEVT 60 Query 66 TSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARV 125 TSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARV Sbjct 61 TSAKEAFADITNTVRARILEIGYDSSDKGFDGATCGVNIGIGAQSPDIAQGVDTAHEARV 120 Query 126 EGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYLRPDG 185 EGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYLRPDG Sbjct 121 EGAADPLDSQGAGDQGLMFGYAINATPELMPLPIALAHRLSRRLTEVRKNGVLPYLRPDG 180 Query 186 KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST 245 KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST Sbjct 181 KTQVTIAYEDNVPVRLDTVVISTQHAADIDLEKTLDPDIREKVLNTVLDDLAHETLDAST 240 Query 246 VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR 305 VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR Sbjct 241 VRVLVNPTGKFVLGGPMGDAGLTGRKIIVDTYGGWARHGGGAFSGKDPSKVDRSAAYAMR 300 Query 306 WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA 365 WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA Sbjct 301 WVAKNVVAAGLAERVEVQVAYAIGKAAPVGLFVETFGTETEDPVKIEKAIGEVFDLRPGA 360 Query 366 IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI 408 IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI Sbjct 361 IIRDLNLLRPIYAPTAAYGHFGRTDVELPWEQLDKVDDLKRAI 403 </span>SUMMER 2017: CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol:
1 ATGTCTGAAAAGGGTCGCCTGTTCAC 26
2 TCGCAGATTTTGTCCGGGTGACCTTCCGTAACAGATTCGCTGGTGAACAGGCGACCCTTT 60
3 CCCGGACAAAATCTGCGACGCAATCTCTGACTCTGTACTGGATGCGCTCCTCGCGGCTGA 60
4 TGACCAGTAGTAACGAGCGTTTCAACCGCAACACGGGAACGTGGGTCAGCCGCGAGGAGC 60
5 ACGCTCGTTACTACTGGTCAGGTACACGTTGTTGGCGAAGTTACCACCTCTGCGAAGGAA 60
6 AGGATACGAGCGCGTACCGTATTCGTGATGTCGGCGAACGCTTCCTTCGCAGAGGTGGTA 60
7 GGTACGCGCTCGTATCCTGGAGATCGGTTACGACTCTTCTGATAAAGGCTTCGATGGTGC 60
8 AGGAGACTGCGCACCGATACCGATATTAACACCACACGTAGCACCATCGAAGCCTTTATC 60
9 CGGTGCGCAGTCTCCTGATATCGCGCAGGGTGTAGATACCGCACACGAAGCGCGTGTTGA 60
10 TTGGTCACCTGCACCCTGAGAGTCGAGCGGGTCTGCCGCACCTTCAACACGCGCTTCGTG 60
11 AGGGTGCAGGTGACCAAGGCCTGATGTTCGGTTATGCCATCAACGCGACCCCGGAACTCA 60
12 GAGACGGCGAGAGAGACGGTGGGCGAGGGCGATCGGCAGCGGCATGAGTTCCGGGGTCGC 60
13 CGTCTCTCTCGCCGTCTCACTGAGGTTCGTAAAAACGGTGTTCTGCCGTACCTGCGTCCT 60
14 GGGACGTTATCTTCGTACGCGATGGTAACCTGGGTTTTACCGTCAGGACGCAGGTACGGC 60
15 GCGTACGAAGATAACGTCCCGGTTCGTCTGGACACTGTTGTTATCTCTACCCAGCACGCA 60
16 CGGATATCTGGGTCCAGAGTTTTTTCCAGATCAATGTCCGCTGCGTGCTGGGTAGAGATA 60
17 ACTCTGGACCCAGATATCCGTGAAAAAGTTCTGAATACCGTCCTGGACGACCTGGCTCAC 60
18 TCGGATTAACCAGAACGCGAACGGTGGAGGCGTCCAGCGTCTCGTGAGCCAGGTCGTCCA 60
19 CGCGTTCTGGTTAATCCGACCGGTAAATTCGTACTGGGTGGTCCGATGGGTGACGCCGGT 60
20 GCCCAACCACCGTAGGTGTCTACAATGATTTTACGGCCGGTGAGACCGGCGTCACCCATC 60
21 ACCTACGGTGGTTGGGCCCGTCACGGTGGCGGTGCGTTTTCCGGCAAAGATCCGTCTAAA 60
22 TCGCAACCCAACGCATCGCATAGGCCGCAGAGCGATCAACTTTAGACGGATCTTTGCCGG 60
23 GATGCGTTGGGTTGCGAAAAATGTTGTTGCTGCGGGCCTCGCTGAACGCGTGGAAGTGCA 60
24 GAACAGACCAACTGGCGCTGCTTTACCAATTGCGTAGGCGACTTGCACTTCCACGCGTTC 60
25 GCGCCAGTTGGTCTGTTCGTCGAAACCTTTGGCACCGAAACCGAAGACCCGGTTAAGATT 60
26 CACCAGGGCGGAGGTCGAAAACTTCACCGATTGCTTTCTCAATCTTAACCGGGTCTTCGG 60
27 GACCTCCGCCCTGGTGCGATCATCCGCGACCTGAACCTGCTGCGCCCAATTTATGCGCCT 60
28 CAGTTCAACGTCCGTACGACCGAAATGACCATAAGCCGCGGTAGGCGCATAAATTGGGCG 60
29 TCGTACGGACGTTGAACTGCCGTGGGAACAGCTGGACAAAGTAGATGATCTCAAGCGTGC 60
30 TTAAATGGCACGCTTGAGATCATCTACT 28
SUMMER 2017 TAIL PRIMER DESIGN
Alexandra B.Forward: 5’ TACTTCCAATCCATGATGTCTGAAAAGGGTCGCC 3'Reverse: 5’ TATCCACCTTTACTGTTAAATGGCACGCTTGAGAT 3’Haoyi W. Forward: 5'TACTTCCAATCCATGATGTCTGAAAAGGGTCGC3'Reverse: 5'TATCCACCTTTACTGTTAAATGGCACGCTTGAGA3'